PubMed Health⌕ Search

Biomedical subjects

T Ueda

Publications and source records attributed to T Ueda.

At least 739 records · Page 41Linked to original sources
PubMed1994

Effect of a peptide from Panax ginseng on the proliferation of baby hamster kidney-21 cells.

An alkaline fraction separated by ion exchange chromatographies from the water extract of Panax ginseng root stimulated the proliferation of baby hamster kidney-21 cells. Separation of the alkaline fraction by MCI-gel CHP 20P column chromatography followed by dialysis provided an active material. By a reversed-phase HPLC the active material was separated into six fractions, and an active colorless compound 1 was obtained from fraction 2 in a pure state. Compound 1 was composed of the following amino acids; Gly, Arg, Glu, Val in a ratio of 1:1:1:1, and caused 20% enhancement of proliferation of BHK-21 cells at a concentration of 3.40 microM. On the basis of physical and spectral data the structure of compound 1 was established as a tetrapeptide, Gly-Arg-gamma-Glu-Val-NH2.

Amino Acid Sequence↗
PubMed1994

The A-domain of beta 2 integrin CR3 (CD11b/CD18) is a receptor for the hookworm-derived neutrophil adhesion inhibitor NIF.

The A-domain is a approximately 200-amino acid peptide present within structurally diverse proadhesive proteins including seven integrins. A recombinant form of the A-domain of beta 2 integrins CR3 and LFA-1 has been recently shown to bind divalent cations and to contain binding sites for protein ligands that play essential roles in leukocyte trafficking to inflammatory sites, phagocytosis and target cell killing. In this report we demonstrate that the neutrophil adhesion inhibitor, NIF produced by the hookworm Ancyclostoma caninium is a selective CD11b A-domain binding protein. NIF bound directly, specifically and with high affinity (Kd of approximately 1 nM) to recombinant CD11b A-domain (r11bA). The binding reaction was characterized by rapid association and very slow dissociation, and was blocked by an anti-r11bA monoclonal antibody. No binding was observed to rCD11aA. The NIF-r11bA interaction required divalent cations, and was absent when the mutant r11bA D140GS/AGA (that lacks divalent cation binding capacity) was used. The NIF binding site in r11bA was mapped to four short peptides, one of which being an iC3b binding site. The interaction of NIF with CR3 in intact cells followed similar binding kinetics to those with r11bA, and occurred with similar affinity in resting and activated human neutrophils, suggesting that the NIF epitope is activation independent. Binding of NIF to CR3 blocked its ability to bind to its ligands iC3b, fibrinogen, and CD54, and inhibited the ability of human neutrophils to ingest serum opsonized particles. NIF thus represents the first example of a disintegrin that targets the integrin A-domain, and is likely to be used by the hookworm to evade the host's inflammatory response. The unique structure of NIF, which lacks a disintegrin motif, emphasizes basic structural differences in antagonists targeting A+ and A- integrins, that should be valuable in drug design efforts aimed at generating novel therapeutics. Identification of the region in NIF mediating A-domain binding should also be useful in this regard, and may, as in the case of disintegrins, unravel a new structural motif with cellular counterparts mediating important physiologic functions.

Amino Acid Sequence↗
PubMed1994

Reduction of disulfide bonds in proteins by 2-aminothiophenol under weakly acidic conditions.

We developed a method for reducing disulfide bonds in proteins under weakly acidic conditions by use of 2-aminothiophenol. The disulfide bonds in hen egg-white lysozyme, ribonuclease A, and soybean trypsin inhibitor were quantitatively reduced by 2-aminothiophenol in phosphate buffer, pH6, containing 8 M Gdn HCl, 1 mM EDTA, and 20% ethanol, for 60 min at 40 degrees C. On analysis of the RP-HPLC patterns of tryptic peptides, which were derived from reduced and S-alkylated lysozyme and ribonuclease A at pH 6, it was confirmed that no side reaction occurred. Moreover, the reduction under weakly acidic conditions was demonstrated to be applicable for the location of such a labile residue as O-acetylated tyrosine.

Aniline Compounds↗
PubMed1994

Template-dependent peptide formation on ribosomes catalyzed by pyridine.

We found that poly(U)-dependent polyphenylalanine synthesis on Escherichia coli ribosomes was extremely accelerated in the presence of a high concentration of pyridine. In this reaction, chemical energy sources, such as ATP and GTP, and soluble protein factors are not required, but the template and ribosomes are essential for the progress of the reaction. The reaction was inhibited by the antibiotics which inhibit the usual bacterial translation process on the ribosomes. These observations clearly demonstrate that the pyridine-catalyzed amino acid condensation reaction proceeds on the ribosome.

Bacterial Proteins↗
PubMed1994

The role of net charge on the renaturation of reduced lysozyme by the sulfhydryl-disulfide interchange reaction.

Reduced and acetylated lysozymes are basic proteins. When their amino groups were variously acetylated and then renatured by sulfhydryl-disulfide (SH-SS) interchange reaction at pH 8.0, the final folding yield decreased as the number of positive charges decreased. The final folding yield of native and Ac1 lysozyme, with one positive charge eliminated, was less sensitive to increasing protein concentration than that of Ac2 lysozyme, where two positive charges had been eliminated. The final folding yield of reduced Ac2 lysozyme increased in the presence of 1 M urea, which reduced the aggregation of unfolded lysozyme. Thus, the aggregation of unfolded lysozymes, which leads to a decrease in the final folding yield, was found to be heavily dependent on their net charges. Moreover, the final folding yield of reduced lysozyme was shown to be increased by use of cystamine as an oxidizing reagent in comparison with 2-hydroxyethyl disulfide or dithiodiglycolic acid. This may support the idea that the final folding yield is influenced by electrostatic interaction between unfolded lysozymes in the early stage of renaturation. In contrast, the concentration dependency of the final folding yield of Ac1 lysozyme was different from those of carboxymethylated His15 and Asp106 lysozymes whose positive net charges were similar to that of Ac1 lysozyme. On the basis of the observations, it is suggested that the formation of the aggregates in the renaturation process might also be affected by the structure of the unfolded state of lysozyme in solution.

Acetylation↗
PubMed1994

Lysozyme requires fluctuation of the active site for the manifestation of activity.

Mutations around His15 which lie far away from the active site, stimulated glycol chitin activity of lysozyme at physiological temperature. Del-Arg14His15 lysozyme, a mutant lysozyme whose Arg14 and His15 were deleted together, and has the highest activity among these mutant lysozymes, had a similar binding ability to a trimer of N-acetyl-glucosamine, a substrate analogue, relative to native lysozyme. This suggests that the increased activity was due to an increased kcat in the catalysis reaction. The H-D exchange rate of the N-1 proton in the Trp63 which is located in the active site cleft, was enhanced in the Del-Arg14His15 lysozyme, while 2-D proton NMR analysis revealed no conformational change around Trp63. We conclude that some sort of fluctuation at the active site might be required for the manifestation of activity. This theory is supported by the finding that the Del-Arg14His15 lysozyme showed a shift in temperature dependency of activity to lower temperatures compared with that of native lysozyme.

Acetylglucosamine↗
PubMed1994

Circumvention of daunorubicin resistance by a new tamoxifen derivative, toremifene, in multidrug-resistant cell line.

The reversing effect of toremifene, a new tamoxifen derivative, on multidrug resistance in a K562 subline and its mechanism were studied. K562 cells were cultured in serially increasing concentrations up to 1.0 microM daunorubicin (DNR), and were found to be 28 times more resistant to DNR in comparison to the parent cells. In the resistant cell line (K562/D1-9), intracellular accumulation of DNR was less than that of the parent cell line, and P-glycoprotein was overexpressed. The resistance was reversed by addition of toremifene in a dose-dependent manner in K562/D1-9, while toremifene had no effect in K562. DNR accumulation was also reversed by toremifene in K562/D1-9, but not in K562. However, there was no significant difference of toremifene retention between K562/D1-9 and K562, and neither verapamil nor DNR increased toremifene accumulation in K562/D1-9. Moreover, toremifene and verapamil did not show an additive effect on intracellular DNR accumulation. These results suggested that the reversing mechanism of toremifene is different from that of verapamil, and this compound could be a good candidate for overcoming multidrug resistance.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1994

Modulation of the effect of 1-beta-D-arabinofuranosylcytosine by 6-mercaptopurine in L1210 cells.

In L1210 cells incubated with 1-beta-D-arabinofuranosylcytosine (ara-C), 6-mercaptopurine (6-MP) significantly potentiated 1-beta-D-arabinofuranosylcytosine 5'-triphosphate (ara-CTP) accumulation and ara-C incorporation into DNA (ara-C/DNA). The cytotoxicity of these two drugs was assessed to be at least additive by clonogenic assay. 1-beta-D-Arabinofuranosyluracil (ara-U) level in a cell suspension was suppressed by 6-MP in a concentration-dependent fashion, though intracellular cytidine deaminase (CDD) activity was not affected by 6-MP. In addition, extracted CDD activity was not directly inhibited by 6-MP or by its intracellular metabolites in vitro. After preincubation in the presence or absence of 6-MP, the cell suspension was fractionated to obtain the spent medium and cell pellet. Then, each fraction was incubated with ara-C. Ara-U formation in the spent medium was found to increase conspicuously in relation to the time of preincubation in the control and it was suppressed by 6-MP pretreatment. Ara-U formation in the cell compartment increased slightly in relation to the time of preincubation in the control and substantially no suppression of ara-U formation was observed in spite of 6-MP pretreatment. In conclusion, intracellularly synthesized CDD was thought to be rapidly shed into the medium and the released CDD could play an important role in ara-C inactivation. 6-MP interrupted some step between synthesis and shedding of CDD, resulting in a decrease of the ara-C deamination in the medium and enhancement of its antileukemic effect.

Animals↗
PubMed1994

Establishment and characterization of a novel myeloid cell line from the bone marrow of a patient with the myelodysplastic syndrome.

A novel long-term cultured myeloid cell line was established from the bone marrow of a patient with myelodysplastic syndrome (MDS). This cell line, designated MDS92, proliferated in the presence of interleukin-3 or granulocyte-macrophage colony-stimulating factor and transiently in the presence of Steel factor, with a tendency for gradual maturation, and formed myeloid colonies in the semi-solid culture condition. In addition, the MDS92 cell line represented rather complicated karyotypic abnormalities including the deletion of fifth and seventh chromosomes and a point mutation at codon 12 of the N-ras oncogene. These characteristics of the MDS92 cell line are exclusively compatible with the property of preleukaemia.

Anemia, Sideroblastic↗
PubMed1994

Evolving experience with continent urinary diversion using the Indiana pouch.

The Indiana continent urinary reservoir procedure for urinary diversion was performed in 46 patients. After the first 8 cases several modifications were made to the Indiana pouch. These modifications include complete detubularization of the colonic segment, construction of the pouch augmented with a U-shaped patch of ileum and, more recently, the use of stapled plication. The mean postoperative follow-up period was 46 mo (range 4.5-74.1 mo). There was 1 perioperative death and this case was excluded from the follow-up study. Late complications related to the urinary reservoir occurred in 12 (26.7%) of the 45 patients. Stone formation was observed in 5 patients. Out of 45 patients, 42 achieved complete urinary continence while 2 suffered mild nocturnal incontinence and 1 had significant leakage. Three had unilateral hydronephrosis due to ureteroanastomotic stricture. Reservoirgraphy demonstrated no reflux into the upper urinary tract in all of the follow-up cases. Loopography of the efferent limb showed that a staple-plicated ileum functions better than a suture-plicated one in terms of reliability of the continence mechanism and ease of catheterization. The Indiana pouch should be considered for any patient requiring cutaneous urinary diversion because it is technically easy to perform and it has a low revision and high continence rate.

Adult↗
PubMed1994

Delayed-type allergenicity of triforine (Saprol).

The delayed-type allergenicity of triforine (Saprol), 1,4-bis (2,2,2-trichloro-1-formamidoethyl) piperazine, was studied. In a mass examination of chrysanthemum growers among whom triforine was commonly used, the highest rate of positive patch test reaction was seen to triforine (17%) among the 7 pesticides and chrysanthemum extracts tested. A higher prevalence rate of work-related skin symptoms was seen in subjects with a positive patch test reaction to triforine (44%) than in those with negative reactions to all allergens tested (15%) (p < 0.05). 12 subjects (67%) with positive patch test reactions to triforine were also positive to dichlorvos (DDVP), with a high kappa coefficient (0.65). The grading of guinea pig maximization test to triforine was grade IV (66%), defined as "strong". Cross-sensitization between triforine and dichlorvos was also shown. The present results confirm that triforine is capable of including delayed-type allergy among chrysanthemum growers and of showing cross-reactivity with dichlorvos.

Adult↗
PubMed1994

Substance P induces inositol 1,4,5-trisphosphate and intracellular free calcium increase in cultured normal human epidermal keratinocytes.

Substance P is a neuropeptide which is present in peripheral C nerve endings and released from them. Free nerve endings of C nerve are present in human epidermis. The effects of substance P on the transmembrane signaling system of pig epidermal sheets were previously reported. In these studies, a small amount of cells other than keratinocytes contaminated the epidermal sheets and the species difference from human was also noticed. Therefore we investigated the effects of substance P on cultured normal human epidermal keratinocytes. Alteration of intracellular free calcium (Ca2+) in single living keratinocytes was studied using an inverted fluorescence microscope and Ca(2+)-sensitive dye, Fura 2-AM. Treatment of normal human epidermal keratinocytes with substance P resulted in an increase in inositol 1,4,5-trisphosphate and in intracellular Ca2+. Substance P inhibited DNA synthesis of the keratinocytes in a dose-dependent manner. These results are consistent with the view that substance P stimulates phosphatidylinositol-4,5-bisphosphate hydrolysis of human keratinocytes, resulting in inositol 1,4,5-trisphosphate-Ca2+ signal.

Calcium↗
PubMed1994

Sequence and spatial requirements for the tissue- and species-independent 3'-end processing mechanism of plant mRNA.

Two cis-regulatory regions are required for efficient mRNA 3'-end processing of the maize 27-kDa zein mRNA: a region containing a duplicated AAUGAA poly(A) signal and a region that is present upstream from it. Strict spatial positioning of these two regions is required for efficient mRNA 3'-end processing. Insertion of a stuffer sequence as short as 17 or 18 bp either between the upstream region and the two AAUGAA motifs or between the two AAUGAA motifs drastically reduced the efficiency of 3'-end processing. Mutational analyses of the nucleotide preference at the fourth position of the AAUGAA motif revealed the preference order G > A >> C or U, suggesting that AAUAAA is neither a defective nor an optimal poly(A) signal for the 27-kDa zein mRNA. As for the 3' control region of the cauliflower mosaic virus (CaMV) transcription unit, the mRNA 3'-end processing mechanism mediated by the 27-kDa zein 3' control sequence is neither tissue nor species specific. The 3' upstream sequence of the 27-kDa zein gene can functionally replace that of the CaMV transcription unit. Conversely, the CaMV upstream sequence can mediate mRNA polyadenylation in the presence of a duplicated 27-kDa zein poly(A) signal. However, instead of the proximal poly(A) signal normally used in the 27-kDa zein mRNA, the distal signal is utilized. These results suggest that a general mechanism controls the 3'-end processing of plant mRNAs and that the cis-regulatory functions mediated by their upstream regions are interchangeable.

Base Sequence↗
PubMed1994

Identification of a transcriptional activator-binding element in the 27-kilodalton zein promoter, the -300 element.

By utilizing a homologous transient-expression system, we have shown that a 58-bp sequence from the gamma-class 27-kDa zein promoter, spanning from -307 to -250 relative to the transcription start site, confers a high level of transcriptional activity on a truncated plant viral promoter. The transcriptional activity mediated by the 58-bp sequence is orientation independent, and it is further enhanced as a result of its multimerization. A similarly high level of transcriptional activity was also observed in protoplasts isolated from leaf tissue-derived maize suspension cells. In vitro binding and DNase I footprinting assays with nuclear protein prepared from cultured endosperm cells revealed the sequence-specific binding of a nuclear factor(s) to a 16-nucleotide sequence present in the 58-bp region. The nuclear factor binding sequence includes the -300 element, a cis-acting element highly conserved among different zein genes and many other cereal storage protein genes. A 23-bp oligonucleotide sequence containing the nuclear factor binding site is sufficient for binding the nuclear factor in vitro. It also confers a high level of transcriptional activity in vivo, but in an orientation-dependent manner. Four nucleotide substitutions in the -300 element drastically reduced binding and transcriptional activation by the nuclear factor. The same nuclear factor is abundant in the developing kernel endosperm and binds to the -300 element region of the 27-kDa or the alpha-class zein promoter. These results suggest that the highly conserved -300 element is involved in the common regulatory mechanisms mediating the coordinated expression of the zein genes.

Base Sequence↗
PubMed1994

Ontogeny and effects of exogenous surfactant treatment on SP-A, SP-B, and SP-C mRNA expression in rabbit lungs.

The aims of the present study were to first characterize the ontogeny of mRNAs encoding the principal surfactant proteins SP-A, SP-B, and SP-C in rabbit lung tissues, followed by an evaluation of the effect of exogenous surfactant administration on endogenous expression of mRNA levels. Lung tissues were collected from New Zealand White rabbits on days 19, 21, 24, 26, 28, 30 of gestation, 1 day and 10 days after birth, and as adults. SP-A, SP-B, and SP-C mRNAs were first detected on gestational days 26, 24, and 19, respectively. The specific levels of these mRNAs generally increased to gestational day 28 and decreased transiently on 1 day of life. Adult values remained lower than those noted at 30 days gestation. For the second study, 29-day-gestation preterm rabbits were given either one or two treatment doses of natural surfactant or an organic solvent extract of natural surfactant, and mRNA levels were measured 10 or 24 h later. No effect of surfactant treatments, type of surfactant, or dosage (1 or 2 treatments) was noted on SP-A, SP-B, or SP-C mRNA expression. These data indicate that the mRNA expression for the three major surfactant proteins was highest late in gestation in developing rabbits and that exogenous surfactant treatment was not associated with significant changes in SP-A, SP-B, or SP-C mRNA levels.

Animals↗